• Title/Summary/Keyword: 16S-rRNA

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Sinomonas terrae sp. nov., Isolated from an Agricultural Soil

  • Hyosun Lee;Ji Yeon Han;Dong-Uk Kim
    • Journal of Microbiology and Biotechnology
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    • v.33 no.7
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    • pp.909-914
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    • 2023
  • While searching for the bacteria which are responsible for degradation of pesticide in soybean field soil, a novel bacterial strain, designated 5-5T, was isolated. The cells of the strain were Gram-staining-positive, aerobic and non-motile rods. Growth occurred at 10-42℃ (optimum, 30℃), pH 5.5-9.0 (optimum, pH 7.0-7.5), and 0-2% (w/v) NaCl (optimum, 1%). The predominant fatty acids were C15:0 anteiso, C17:0 anteiso, and summed feature 8 (C18:1 ω7c and/or C18:1 ω6c). The predominant menaquinone was MK-9 (H2). Diphosphatidylglycerol, glycolipids, phosphatidylinositol, and phosphatidylglycerol were the major polar lipids. Phylogenetic analysis of 16S rRNA gene sequences indicated that strain 5-5T is a member of the genus Sinomonas and its closest relative is Sinomonas humi MUSC 117T, sharing a genetic similarity of 98.4%. The draft genome of strain 5-5T was 4,727,205 bp long with an N50 contig of 4,464,284 bp. Genomic DNA G+C content of strain 5-5T was68.0 mol%. The average nucleotide identity (ANI) values between strain 5-5T and its closest strains S. humi MUSC 117T and S. susongensis A31T were 87.0, and 84.3 % respectively. In silico DNA-DNA hybridization values between strain 5-5T and its closest strains S. humi MUSC 117T and S. susongensis A31T were 32.5% and 27.9% respectively. Based on the ANI and in silico DNA-DNA hybridization analyses, the 5-5T strain was considered as novel species belonging to the genus Sinomonas. On the basis of the results from phenotypic, genotypic and chemotaxonomic analyses, strain 5-5T represents a novel speciesof the genus Sinomonas, for which the name Sinomonas terrae sp. nov. is proposed. The type strain is 5-5T (=KCTC 49650T =NBRC 115790T).

Effect of Extracts and Bacteria from Korean Fermented Foods on the Control of Sesame Seed-Borne Fungal Diseases (발효식품 추출물과 미생물을 활용한 참깨 종자전염성 병 방제)

  • Kim, Yong-Ki;Hong, Sung-Jun;Shim, Chang-Ki;Kim, Min-Jeong;Park, Jong-Ho;Han, Eun-Jung;Park, Jong-Won;Park, So-Hyang;Jee, Hyeong-Jin;Kim, Seok-Cheol
    • Research in Plant Disease
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    • v.21 no.4
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    • pp.297-308
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    • 2015
  • In order to control seed-borne diseases, we obtained extracts from commercial fermented food products of Kimchi, Gochujang, Doenjang, Ganjang and Makgeolli and their suppressive effects against seed-borne diseases were studied. In addition, the suppressive effects of bacterial strains isolated from the fermented foods were screened in vitro and in vivo. Among fifty food extracts, twenty food-extracts suppressed more than 92% incidence of seedling rots in vitro and seven food extracts increased 58.3-66.8% of healthy seedling in the greenhouse. Among 218 isolates from the fermented foods, 29 isolates showing high antifungal activity against seven seed-borne fungal pathogens were selected. Among 29 isolates, 13 isolates significantly reduced seedling rot and increased healthy seedlings. Sixteen isolates with high antifungal activity and suppressive effect against sesame seedling rots were identified by 16S rRNA sequencing. Fourteen of sixteen isolates were identified as Bacillus spp. and the other two isolates from Makgeolli were identified as Saccharomyces cerevisiae. It was confirmed that B. amyloliquifaciens was majority in the effective bacterial population of Korean fermented foods. In addition, when the bioformulations of the two selected effective microorganisms, B. amyloliquifacien Gcj2-1 and B. amyloliquifacien Gcj3-1, were prepared in powder forms using bentonite, kaolin, talc and zeolite, talc- and kaolin-bioformulation showed high control efficacy against sesame seed-borne disease, followed by zeolite-bioformulation. Meanwhile control efficacy of each bentonite-bioformulation of B. amyloliquifacien Gcj2-1 and B. amyloliquifacien Gcj3-1 was lower than that of bacterial suspension of them. It was found that the selected effective microorganisms from Korean fermented foods were effective for controlling seed-borne diseases of sesame in vitro and in the greenhouse. We think that Korean fermented food extracts and useful microorganisms isolated from the extract can be used as bio-control agents for suppressing sesame seed-borne diseases based on above described results.

The effect of antagonists produced by Paenibacillus polymyxa CK-1 on the growth of Trichoderma sp. (Paenibacillus polymyxa CK-1이 생산한 길항물질이 Trichoderma sp. 생육에 미치는 영향)

  • Lee, Sang-Won;Choi, Jin-Sang;Kim, Chul-Ho
    • Journal of Mushroom
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    • v.12 no.3
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    • pp.201-208
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    • 2014
  • The separation of the bacteria inhibiting Trichoderma sp. mold, the strain causing blue mold disease that occurs frequently when cultivating mushroom while carrying out the efficient fermentation of mushroom medium, from the growth was done. In about 200 strains isolated primarily from fungus garden samples, 6 strains were secondly isolated, which had fast growth rates and a clear zone on the plate medium of SM, AM, and CM. Among the 6 strains isolated, the C-1 strain showed high enzymatic activity of cellulase, amylase, and protease, and strong antibacterial activity for the T. virens and T. harzianum, selected finally. The selected C-1 strain was identified as Paenibacillus polymyxaby the result of the identification by Bergey's Manual of Systematic Bacteriology and the analysis of the nucleotide sequence of 16S rRNA, and named as P. polymyxa CK-1. In reviewing the growth conditions of the P. polymyxa CK-1 strain, the optimum cultivation temperature was $45^{\circ}C$, and the optimum pH for growth was in the range of 6.0~7.0. Appropriate incubation time of P. polymyxa CK-1 for the growth inhibition of the fungus T. virens and T. harzianum was 22 to 36 hours. And the fungal growth was not observed, even when leaving two molds inoculated on each petri dishes, which were treated with 24 hour culture solution of P. polymyxa CK-1 strain for 10 days. As a result of studying the thermal stability of the antagonists produced by the P. polymyxa CK-1 strain, no mycelial growth of the two fungi was observed in the test group treated for 20 minutes at $60^{\circ}C$ and $100^{\circ}C$, but mycelial growth was slightly observed in the test group treated for 20 minutes at $121^{\circ}C$. As aresult of reviewing the impact of the P. polymyxa CK-1 culture medium on mushroom mycelial growth, it showed no effect on a variety of mushroom mycelial growth including enoki mushroom and shiitake mushroom.

Deacidification Effect of Campbell Early Must through Carbonic-Maceration Treatment: Isolation and Properties of the Bacteria Associated with Deacidification (Carbonic Maceration처리에 의한 Campbell Early 발효액의 감산 효과: 감산 관련 미생물의 분리 및 특성)

  • Chang, Eun-Ha;Jeong, Seok-Tae;Jeong, Sung-Min;Lim, Byung-Sun;Noh, Jung-Ho;Park, Kyo-Sun;Park, Seo-Jun;Choi, Jong-Uck
    • Food Science and Preservation
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    • v.18 no.6
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    • pp.973-979
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    • 2011
  • The grape cultivar Campbell Early has high levels of malic acid as well as tartaric acid. The high concentration of total acid in the Campbell Early wine is a critical aspect of the wine's sensory characteristics. To prevent the deterioration of the wine's quality, which is caused by the strong sour taste derived from the raw material in wine making, the deacidification factor was investigated via carbonic maceration under different temperature conditions, especially in the presence or absence of malolactic bacteria. Based on the results of the presence test of the malolactic bacteria during carbonic-maceration treatment, Lactobacillus brevis, Lactobacillus plantarum, and Streptococcus thermophilus were characterized morphologically and were identified via biochemical tests and 16S-rRNA-gene-sequencing analysis. The isolated strains were found not to consume malic acid and to produce lactic acid. Moreover, these strains were consumed as soluble solids. The isolated strains are popularly known as lactic-acid bacteria and should have produced lactic acid from glucose. The Oenococcus oeni of the malolactic bacteria was not isolated. These results showed that the isolated strains are not deacidified during carbonic-maceration treatment.

The Fermentative Hydrogen Production in Trickling Bed Biofilter Filled with Hydrophilic-and Hydrophobic-Media (소수성 및 친수성 담체를 이용한 Trickling Bed Biofilter의 생물학적 수소생산)

  • Jeon, Byung-Seung;Lee, Sun-Mi;Kim, Yong-Hwan;Gu, Man-Bock;Chae, Hee-Jeong;Sang, Byoung-In
    • Transactions of the Korean hydrogen and new energy society
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    • v.17 no.4
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    • pp.379-388
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    • 2006
  • Two mesophilic trickling bed bioreactors filled with two different types of media, hydrophilic- and hydrophobic-cubes, were designed and conducted for hydrogen production under the anaerobic fermentation of sucrose. Each bioreactor consisted of the column packed with polymeric cubes and inoculated with heat-treated sludge obtained from anaerobic digestion tank. A defined medium containing sucrose was fed by the different hydraulic retention time(HRT), and recycle rate. Hydrogen concentrations in gas-phase were constant, averaging 40% of biogas throughout the operation. Hydrogen production rate was increased till $10.5\;L{\cdot}h^{-1}{\cdot}L^{-1}$ of bioreactor when influent sucrose concentrations and recycle rates were varied. At the same time, the hydrogen production rate with hydrophobic media application was higher than its hydrophilic media application. No methane was detected when the reactor was under a normal operation. The major fermentation by-products in the liquid effluent of the both trickling biofilters were acetate, butyrate and lactate. In order to run in the long term operation of both reactor filled with hydrophilic and hydrophobic media, biofilm accumulation on hydrophilic media and biogas produced should be controlled through some process such as periodical backwashing or gas-purging. Four sample were collected from each reactor on the opposite hydrogen production rate, and their bacterial communities were compared by terminal restriction fragment length polymorphism (T-RFLP) analysis of PCR products generated using bacterial 16s rRNA gene primers (8f and 926r). It was expressed a marked difference in bacterial communities of both reactors. The trickling bed bioreactor with hydrophobic media demonstrates the feasibility of the process to produce hydrogen gas. A likely application of this reactor technology can be hydrogen gas recovery from pre-treatment of high carbohydrate-containing wastewaters.

Antifouling Activity towards Mussel by Small-Molecule Compounds from a Strain of Vibrio alginolyticus Bacterium Associated with Sea Anemone Haliplanella sp.

  • Wang, Xiang;Huang, Yanqiu;Sheng, Yanqing;Su, Pei;Qiu, Yan;Ke, Caihuan;Feng, Danqing
    • Journal of Microbiology and Biotechnology
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    • v.27 no.3
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    • pp.460-470
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    • 2017
  • Mussels are major fouling organisms causing serious technical and economic problems. In this study, antifouling activity towards mussel was found in three compounds isolated from a marine bacterium associated with the sea anemone Haliplanella sp. This bacterial strain, called PE2, was identified as Vibrio alginolyticus using morphology, biochemical tests, and phylogenetic analysis based on sequences of 16S rRNA and four housekeeping genes (rpoD, gyrB, rctB, and toxR). Three small-molecule compounds (indole, 3-formylindole, and cyclo (Pro-Leu)) were purified from the ethyl acetate extract of V. alginolyticus PE2 using column chromatography techniques. They all significantly inhibited byssal thread production of the green mussel Perna viridis, with $EC_{50}$ values of $24.45{\mu}g/ml$ for indole, $50.07{\mu}g/ml$ for 3-formylindole, and $49.24{\mu}g/ml$ for cyclo (Pro-Leu). Previous research on the antifouling activity of metabolites from marine bacteria towards mussels is scarce. Indole, 3-formylindole and cyclo (Pro-Leu) also exhibited antifouling activity against settlement of the barnacle Balanus albicostatus ($EC_{50}$ values of 8.84, 0.43, and $11.35{\mu}g/ml$, respectively) and the marine bacterium Pseudomonas sp. ($EC_{50}$ values of 42.68, 69.68, and $39.05{\mu}g/ml$, respectively). These results suggested that the three compounds are potentially useful for environmentally friendly mussel control and/or the development of new antifouling additives that are effective against several biofoulers.

Enzyme Production of A Protease-producing Strain, Bacillus sp SH-8 Isolated from Insect-eating Plant (식충식물로부터 Protease를 생산하는 Bacillus sp. SH-8의 분리와 효소 생산성)

  • Yoon, Ki-Hong;Lee, Mi-Sung;Park, Bueng-Wan;Park, Yong-Ha;Kim, Hong-Ik;Kim, Jeong-Hyeon;Kim, Moon-Sook
    • Microbiology and Biotechnology Letters
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    • v.34 no.4
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    • pp.323-328
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    • 2006
  • A bacterium producing the extracellular protease was isolated from insect-eating plant and has been identified as a member of the genus Bacillus based on partial 165 rRNA sequences. In order to develop the medium composition, effects of ingredients including nitrogen sources, carbon source, metal ions and phosphate were examined for protease production of the isolate, SH-8. Soluble starch increased the protease productivity, while glucose repressed it. Yeast extract was effective nitrogen source for enzyme production, but the pretense production of Bacillus sp. SH-8 was reduced by large amount of yeast extract. The calcium was found to induce pretense activity as well as protease productivity. However, cell growth and enzyme production was completely inhibited by divalent ions such as $Zn^{2+}$, $Cu^{2+}$, $Co^{2+}$ and $Mn^{2+}$. The maximum protease productivity was reached 435 unit/ml in the optimized medium consisting of soluble starch (2%), yeast extract (0.3%), $CaCl_2$ (0.3%), $K_2HPO_4$ (0.01%) and $KH_2PO_4$ (0.01%). The pretense activity of culture filtrate was dramatically decreased after incubation for 26 h.

Selection of Lactic Acid Bacteria with Antibacterial Activity for Extension of Kimchi Shelf-life (김치의 저장성 향상을 위한 항균활성 우수 유산균 선발)

  • Choi, Hak-Jong;Kim, Yu Jin;Lee, Na Ra;Park, Hae Woong;Jang, Ja Young;Park, Sung-Hee;Kang, Miran;Kim, Hyun Ju;Lee, Jong-Hee;Lee, Jong-Hoon;Pyun, Yu-Ryang;Kim, Tae-Woon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.2
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    • pp.328-332
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    • 2014
  • A survey was conducted on the isolation of lactic acid bacteria with antibacterial activity to extend kimchi shelf-life. Antibacterial activity was tested against bacteria associated with acidification of kimchi, including Lactobacillus plantarum, Pediococcus pentosaceus, and Lactobacillus sakei, using agar-well diffusion assay. Two isolates from kimchi were identified as Lactococcus lactis subsp. lactis and Lactobacillus brevis by 16S rRNA gene sequence analysis and API 50 CHL assay, and they showed antibacterial effects against indicator strains. The isolates displayed acid tolerance at pH 3.5, salt tolerance in 5% NaCl, and growth at $4^{\circ}C$. These result imply that the selected strains might be used to extend kimchi shelf-life as a potential starter.

Comparative analysis of the microbial communities in raw milk produced in different regions of Korea

  • Kim, In Seon;Hur, Yoo Kyung;Kim, Eun Ji;Ahn, Young-Tae;Kim, Jong Geun;Choi, Yun-Jaie;Huh, Chul Sung
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.11
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    • pp.1643-1650
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    • 2017
  • Objective: The control of psychrotrophic bacteria causing milk spoilage and illness due to toxic compounds is an important issue in the dairy industry. In South Korea, Gangwon-do province is one of the coldest terrains in which eighty percent of the area is mountainous regions, and mainly plays an important role in the agriculture and dairy industries. The purposes of this study were to analyze the indigenous microbiota of raw milk in Gangwon-do and accurately investigate a putative microbial group causing deterioration in milk quality. Methods: We collected raw milk from the bulk tank of 18 dairy farms in the Hoengseong and Pyeongchang regions of Gangwon-do. Milk components were analyzed and the number of viable bacteria was confirmed. The V3 and V4 regions of 16S rRNA gene were amplified and sequenced on an Illumina Miseq platform. Sequences were then assigned to operational taxonomic units, followed by the selection of representative sequences using the QIIME software package. Results: The milk samples from Pyeongchang were higher in fat, protein, lactose, total solid, and solid non-fat, and bacterial cell counts were observed only for the Hoengseong samples. The phylum Proteobacteria was detected most frequently in both the Hoengseong and Pyeongchang samples, followed by the phyla Firmicutes and Actinobacteria. Notably, Corynebacterium, Pediococcus, Macrococcus, and Acinetobacter were significantly different from two regions. Conclusion: Although the predominant phylum in raw milk is same, the abundances of major genera in milk samples were different between Hoengseong and Pyeongchang. We assumed that these differences are caused by regional dissimilar farming environments such as soil, forage, and dairy farming equipment so that the quality of milk raw milk from Pyeongchang is higher than that of Hoengseong. These results could provide the crucial information for identifying the microbiota in raw milk of South Korea.

Optimization of Indole-3-Acetic production by phosphate solubilization bacteria isolated from waste mushroom bed of Agaricus bisporus

  • Walpola, Buddhi Charana;Noh, Jae-Geun;Kim, Chan Kyem;Kyung, Ki-Cheon;Kong, Won-Sik;Yoon, Min-Ho
    • Journal of Mushroom
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    • v.11 no.2
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    • pp.53-62
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    • 2013
  • A total of 35 phosphate solubilizing bacterial strains were isolated from waste mushroom bed of Agaricus bisporus in Buyeo-Gun, Chungnam and screened for the production of indole acetic acid (IAA). The best IAA producing strain was identified as Pantoea rodasii using 16S rRNA analysis. In addition to the IAA production, this strain could act as an efficient phosphate solubilizer (1100 ${\mu}g$ $ml^{-1}$ after 5 days of incubation) also. The selected strain was cultured under different conditions in order to assess the optimum conditions for maximum IAA production. The nutrient broth (NB) medium was recorded as the best medium, where the maximum IAA production (229 ${\mu}g$ $ml^{-1}$) was recorded at the start of stationary phase (12 hours after inoculation) of the bacteria growth. The performance of the strain was found to be maximum at the temperature of $30^{\circ}C$ followed by $25^{\circ}C$. IAA production was found to be increased with increasing tryptophan concentration (from 0.1 to 0.6%), however beyond this limit, a slight reduction in IAA production was observed. The strains' ability to produce IAA was further confirmed by extraction of crude IAA and subsequent TLC analysis. A specific spot from the extracted IAA preparation was found corresponding with the standard spot of IAA with same $R_f$ value. The results of HPLC analysis conducted in identifying and quantifying the IAA production more precisely, are in agreement with the results of the assessment done with colorimetric method. As revealed by the results of the pot experiment, the isolated strain could significantly enhance the growth (as measured by shoot and root growth) of mung bean plants compared to that of non-inoculated plants. Therefore it can be concluded that the present strain, Pantoea rodasii has great potential to be used as bio-inoculants.